Method for separating and purifying polyhydroxyalkanoate from interiors of microbial cells

Abstract

The invention provides a method for separating and purifying polyhydroxyalkanoate from the interiors of microbial cells. The method includes the following steps: after a microbial strain producing polyhydroxyalkanoate is fermented, the bacterial cells in fermentation broth are collected; in an ultrasonic generator, the bacterial cells are mixed with organic solvent, the dosage of the organic solvent is 10 to 30 times as much as the bacterial cells, and after 20 to 70 minutes of ultrasonic vibration under 40 DEG C to 80 DEG C, solid-liquid mixture is obtained; the solid-liquid mixture is separated centrifugally or by filtration, so that a liquid phase containing polyhydroxyalkanoate is obtained; the liquid phase containing polyhydroxyalkanoate is cooled to 0 DEG C to 10 DEG C, the temperature is kept for 10 to 30 minutes, so that turbid liquid which is formed after polyhydroxyalkanoate crystals are separated out is obtained; the turbid liquid is separated centrifugally or by filtration, so that solid polyhydroxyalkanoate is obtained; and after the solid polyhydroxyalkanoate is dried, finished polyhydroxyalkanoate product is obtained. The method has the advantages of simple process, low energy consumption, low production cost and high product extraction rate, and is easy to operate.

Description

technical field

[0001] The invention relates to the technical field of separation and purification of downstream products of bioengineering, in particular to a method for separating and purifying polyhydroxyalkanoate from microbial cells. Background technique

[0002] Polyhydroxyalkanoate (Polyhydroxyalkanoate, PHAs) is a biopolymer material that has been developed rapidly in the past two decades. It plays an important role in environmental protection and medical applications. It has become a direction for the production, development and application of new materials. It is one of the most active hotspots in the field of biomaterials. Polyhydroxyalkanoate is a type of intracellular thermoplastic polyester synthesized by microorganisms to store excess carbon sources in the form of carbon sources and energy sources, and the cumulative amount can account for 30% - 80% of the dry weight of cells. For PHAs, when the number of carbon atoms in the polymerized monomer is 3-5, the re...

Examples

Embodiment Construction

[0018] The present invention will be further described in detail below in conjunction with the examples, but not limited thereto.

[0019] Now use from Ralstonia eutropha Synthesis (for the specific synthetic method, refer to the article Optimization of Propionic Acid Feeding for Production of Poly (3-hydroxybutyrate- co -3-hydroxyvalerate) in Fed-Batch Culture of Ralstonia eutropha . Chinese Journal of Chemical Engineering. 11(2):220-223) Separating and purifying polyhydroxyalkanoate from bacterial cells containing polyhydroxyalkanoate as an example, the present invention is described in detail, and the specific steps are as follows:

[0020] (1) After fermenting the microbial strain producing polyhydroxyalkanoate, centrifuge the fermentation broth, discard the supernatant to obtain polyhydroxyalkanoate-containing Ralstonia eutropha wet cells. After weighing the fermentation Ralstonia eutropha Wet cells 50 grams.

[0021] (2) In the ultrasonic generator (300W), 50 grams...

technical field

[0001] The invention relates to the technical field of separation and purification of downstream products of bioengineering, in particular to a method for separating and purifying polyhydroxyalkanoate from microbial cells. Background technique

[0002] Polyhydroxyalkanoate (Polyhydroxyalkanoate, PHAs) is a biopolymer material that has been developed rapidly in the past two decades. It plays an important role in environmental protection and medical applications. It has become a direction for the production, development and application of new materials. It is one of the most active hotspots in the field of biomaterials. Polyhydroxyalkanoate is a type of intracellular thermoplastic polyester synthesized by microorganisms to store excess carbon sources in the form of carbon sources and energy sources, and the cumulative amount can account for 30% - 80% of the dry weight of cells. For PHAs, when the number of carbon atoms in the polymerized monomer is 3-5, the re...

Examples

Embodiment Construction

[0018] The present invention will be further described in detail below in conjunction with the examples, but not limited thereto.

[0019] Now use from Ralstonia eutropha Synthesis (for the specific synthetic method, refer to the article Optimization of Propionic Acid Feeding for Production of Poly (3-hydroxybutyrate- co -3-hydroxyvalerate) in Fed-Batch Culture of Ralstonia eutropha . Chinese Journal of Chemical Engineering. 11(2):220-223) Separating and purifying polyhydroxyalkanoate from bacterial cells containing polyhydroxyalkanoate as an example, the present invention is described in detail, and the specific steps are as follows:

[0020] (1) After fermenting the microbial strain producing polyhydroxyalkanoate, centrifuge the fermentation broth, discard the supernatant to obtain polyhydroxyalkanoate-containing Ralstonia eutropha wet cells. After weighing the fermentation Ralstonia eutropha Wet cells 50 grams.

[0021] (2) In the ultrasonic generator (300W), 50 grams...